Biology subjects

Serralbo, O.

Publications and source records attributed to Serralbo, O..

4 recordsLinked to original sources

bioRxiv2026

A Cre-responsive reporter quail for flexible genetic lineage tracing

Genetic lineage tracing is a powerful approach for investigating cell fate and tissue development, but versatile Cre/loxP-based tools remain limited in avian models. Here, we generated and characterised a Cre-responsive transgenic quail line that enables permanent genetic labelling of embryonic cell populations. The reporter line ubiquitously expresses nuclear-localised mCherry, which is excised following Cre-mediated recombination, enabling expression of nuclear-localised EGFP. We demonstrate that this reporter can be coupled to multiple Cre delivery strategies, providing flexible spatial and temporal control of lineage labelling. Ubiquitous or cell-type-specific Cre expression induces recombination and stable EGFP labelling that is maintained during clonal expansion. Optogenetic activation of a split Cre system enables highly spatially and temporally restricted recombination. Furthermore, lipid nanoparticle-mediated delivery of Cre mRNA enables efficient recombination in distinct embryonic cell populations, including mesodermal, neuroepithelial, neural crest, endothelial and circulating cells. This transgenic reporter quail provides a flexible platform for permanent lineage labelling without the need to generate tissue-specific Cre transgenic lines and can be readily combined with diverse Cre delivery strategies to investigate cell fate, lineage and morphogenesis in vivo.

developmental biology↗

bioRxiv2023

The Lifeact-EGFP Quail: A New Avian Model for Studying Actin Dynamics In Vivo

Here we report the generation of a transgenic Lifeact-EGFP quail line for the investigation of actin organization and dynamics during morphogenesis in vivo. This transgenic avian line allows for the high-resolution visualization of actin structures within the living embryo, from the subcellular filaments that guide cell shape to the supracellular assemblies that coordinate movements across tissues. The unique suitability of avian embryos to live imaging facilitates the investigation of previously intractable processes during embryogenesis. Using high-resolution live imaging approaches, we present the dynamic behaviours and morphologies of cellular protrusions in different tissue contexts. Furthermore, through the integration of live imaging with computational segmentation, we reveal the dynamics of cells undergoing apical constriction and the emergence of large-scale actin structures such as supracellular cables and rosettes within the neuroepithelium. These findings not only enhance our understanding of tissue morphogenesis but also demonstrate the utility of the Lifeact-EGFP transgenic quail as a new model system for live in vivo investigations of the actin cytoskeleton.

developmental biology↗

bioRxiv2021

TopFlash transgenic quail reveals dynamic TCF/beta-catenin signaling during avian embryonic development

The Wnt/{beta}-catenin signaling pathway is highly conserved throughout evolution and it plays crucial roles in several developmental and pathological processes. Wnt ligands can act at a considerable distance from their sources and it is therefore necessary to examine not only the Wnt-producing but also the Wnt-receiving cells and tissues to fully appreciate the many functions of this pathway. To monitor Wnt activity, multiple tools have been designed which consist of multimerized Wnt signaling response elements (TCF/LEF binding sites) driving the expression of fluorescent reporter proteins (e.g. GFP, RFP) or of LacZ. The high stability of those reporters leads to a considerable accumulation in cells activating the pathway, thereby making them easily detectable. However, this makes them unsuitable to follow temporal changes of the pathways activity during dynamic biological events. Even though fluorescent transcriptional reporters can be destabilized to shorten their half-lives, this dramatically reduces signal intensities, particularly when applied in vivo. To alleviate these issues, we developed two transgenic quail lines in which high copy number (12x or 16x) of the TCF/LEF binding sites drive the expression of destabilized GFP variants. Translational enhancer sequences derived from viral mRNAs were used to increase signal intensity and specificity. This resulted in transgenic lines efficient for the characterisation of TCF/{beta}-catenin transcriptional dynamic activities during embryogenesis, including using in vivo imaging. Our analyses demonstrate the use of this transcriptional reporter to unveil novel aspects of Wnt signaling, thus opening new routes of investigation into the role of this pathway during amniote embryonic development.

developmental biology↗

bioRxiv2019

Generation of transgenic quails by in vivo transfection of primordial germ cells

During development, dynamic changes in tissue shapes known as morphogenesis result from the exquisite orchestration of signalling pathways, cell-cell interactions, cell divisions, and coordinated movements. How cells within embryos adopt a vast array of cell fates and tissue shapes in such an ever-changing environment has fascinated scientists for generations, yet the ability to observe and characterize those rapid changes has proven technically challenging in higher vertebrates. The japanese quail (Coturnix coturnix japonica) is an attractive model where basic rules driving morphogenesis in amniotes can be deciphered using genetic approaches. Similar to the more popular chicken model, the quail embryo is easily accessible to a wide range of manipulations and live imaging. A decisive asset of quail over chicken is a much shorter life cycle, which makes its use as genetic model for basic research extremely appealing. To date, all existing transgenic quail lines were generated using replication-deficient lentiviruses, but diverse limitations of this approach have hindered the widespread expansion of such technology. Here, we successfully used a plasmid-based in vivo transfection of quail primordial germ cells (PGCs) to generate a number of transgenic quail lines over a short period of time. The plasmid-based approach is simple, efficient and it allows using the infinite variety of genome engineering approaches developed in other models, such as strategies to facilitate transgenic bird selection, shown here. This major technological development completes the vast panel of techniques applicable to the avian model, making it one of the most versatile experimental systems available.

developmental biology↗